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5’-methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTAN) plays a critical role in diverse pathways in bacterialcells such as biological methylation, polyamine biosynthesis, methionine recycling, and bacterial quorum sensing. It hasbeen known that MtaN catalyzes the hydrolysis of N-ribosidic bond of adenosine-based substrates such as S-adenosyl-Lhomocysteine(SAH), S-methyl-5’-thioadenosine (MTA) and 5’-deoxyadenosine (5’-DOA). In Aeromonas hydrophila, thereare two MtnN subfamily proteins: MtaN-1, a periplasmic protein with an N-terminal signal peptide; and MtaN-2, a cytosolicprotein. In this study, MtaN-2 from A. hydrophila was successfully expressed and purified using Ni-NTA affinity, Q anionexchange,and gel-filtration chromatography. We first crystallized apo MtaN-2 but it diffracted to a low resolution of 5.1Å. New crystals suitable for diffraction were obtained by adding 2 mM adenosine, a substrate analog of MtaN-2 duringpurification process and the crystals diffracted to the resolution of 2.0 Å. The crystals belong to the trigonal space groupP31 or P32, with unit-cell parameters of a = b = 74.94 Å and c = 185.21 Å. The asymmetric unit contains four complexes ofMtaN-2 with hydrolysis products of adenosine.
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참고문헌 (18)
참고문헌 신청
Adams, P. D / 2002 / PHENIX : building new software for automated crystallographic structure determination / Acta Crystallogr D Biol Crystallogr 58:1948~1954
Allart, B / 1998 / The catalytic mechanism of adenosylhomocysteine/methylthioadenosine nucleosidase from Escherichia coli—chemical evidence for a transition state with a substantial oxocarbenium character / Eur J Biochem 256:155~162
Beeston, A. L / 2002 / Pfs-dependent regulation of autoinducer 2 production in Salmonella enterica serovar Typhimurium / J Bacteriol 184:3450~3456
Borchardt, R. T / 1986 / Biological Methylation and Drug Design-Experimental and Clinical Roles of S-Adenosyl-methionine / Humana Press :227~238
Challand, M. R / 2009 / Product inhibition in the radical S-adenosylmethionine family / FEBS Lett 583:1358~1362
Allart, B / 1998 / The catalytic mechanism of adenosylhomocysteine/methylthioadenosine nucleosidase from Escherichia coli—chemical evidence for a transition state with a substantial oxocarbenium character / Eur J Biochem 256:155~162
Beeston, A. L / 2002 / Pfs-dependent regulation of autoinducer 2 production in Salmonella enterica serovar Typhimurium / J Bacteriol 184:3450~3456
Borchardt, R. T / 1986 / Biological Methylation and Drug Design-Experimental and Clinical Roles of S-Adenosyl-methionine / Humana Press :227~238
Challand, M. R / 2009 / Product inhibition in the radical S-adenosylmethionine family / FEBS Lett 583:1358~1362
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